Promotion of the population growth of pinewood nematode in 4-month-old Japanese black pine seedlings by pretreatment with simulated acid rain

Simulated acid rain (SAR) at three pH levels (pH 4, 3, 2) was applied to only the top or both the top and roots of 4-month-old Japanese black pine seedlings repeatedly for two months. Then the seedlings were inoculated with a virulent isolate (S10) of pinewood nematode,Bursaphelenchus xylophilus. The exposure to SAR at any level did not kill the seedlings. When inoculated with nematodes, however, the nematode population build-up was significantly higher in the seedlings pretreated with SAR at any level than in the control at the 7th and 17th day after inoculation, and subsequent disease symptom development was also significantly accelerated by the exposure to SAR at pH 2 and 3. This result indicates that even acid rain at pH 4 has the potential for promoting population growth of pinewood nematodes in 4-month-old Japanese black pine seedlings.     

米老排生长与土壤条件的关系及防火林带生态效益的研究

对永春碧卿国有林场山地防火林带的调查研究表明：山地防火林带土壤肥力可分为３个等级,林木生长与土壤肥力等级呈正相关。土壤容重、土壤水分、土壤有机质、全氮、速效钾等几个指标可以综合反映山地防火林带（路）的土壤肥力水平。米老排防火林带具有明显的经济效益和生态效益。     

Retardation of pine wilt disease symptom development in Japanese black pine seedlings exposed to simulated acid rain and inoculated withBursaphelenchus xylophilus

Three- or 4-year-old Japanese black pine seedlings were exposed to simulated acid rain (SAR) at pH 3 for two months, then inoculated with a virulent isolate (S 10) of pinewood nematodes. The experiments were repeated three times in 1996–1998. The exposure to SAR killed no seedlings, and retarded the development of disease symptoms in the seedlings inoculated with nematodes. In the experiments in 1996 and 1997, however, cessation of resin exudation, and primary decrease in the xylem conductivity occurred earlier in the seedlings exposed to SAR than in those exposed to tap water as a control. These results imply that acid rain at pH 3 influenced an increase in some resistance of Japanese black pine seedlings to pinewood nematode, and that this improved resistance could potentially overcome damage caused by acid rain. The exposure to SAR did not have any significant effect on the water relations of the seedlings itself, suggesting that retardation of the symptom development after the exposure to SAR could not be attributed to the improvement in the water relations. This work was supported in part by a Grants-in-Aid for Scientific Research from the Japan Forest Technical Association (JAFTA).     

The role of litter and humus layer as a mortality factor of seedlings of Japanese red pine,Pinus densiflora Sieb. et Zucc.

The process of decrease in seedlings of Japanese red pine (Pinus densiflora Sieb.et Zucc.) and the effect of litter and humus layers on the mortality were studied in their stand and adjacent bare land. After the rainy season, only small number of individuals died in the bare land but more seedlings died in the forest. The number of seedlings killed by drought was greater in the forest than in the bare land. The population of naturally occurring current year seedlings was larger in the forest than in the bare land, but that of 1- to 5-year-old seedlings was smaller. Severe summer drought exceeding the permanent wilting point was observed only in the forest F-H horizon where greater part of seedling roots distributed, this may be one of major mortality factors. Inhibitory substances to seedling growth in fresh fallen pine needles were also suggested.     

The effects of long‐term exposure to simulated acid rain on the development of pine wilt disease caused by Bursaphelenchus xylophilus

To elucidate the synergetic effects of acid rain on the development of pine wilt disease, we measured the sap flow rate in the stems and the chlorophyll content in the needles of 10‐year‐old Japanese black pine trees, Pinus thunbergii and 12‐year‐old Japanese red pine trees, Pinus densiflora, after exposure to simulated acid rain (SAR, pH 3) or tap water (TW, pH 6.3) as a control. The heat pulse method was used for the estimation of the sap flow rate. No apparent difference was found in the sap flow rate between the trees exposed to SAR and TW, but the chlorophyll content of needles at the end of the treatment was significantly higher in the trees exposed to SAR than in those exposed to TW. When the pinewood nematode, Bursaphelenchus xylophilus, the causal agent of pine wilt disease, was inoculated onto the Japanese black pines that had been exposed to SAR repeatedly for 1 year, the period to death was shortened. Japanese red pines that had been exposed to SAR for 2 years, however, did not show any development of symptoms after the nematode inoculation, suggesting that acid rain only affects pine wilt disease slightly, if at all.     

Effects of inoculum density of pinewood nematode on the development of pine wilt disease in Japanese black pine seedlings pretreated with simulated acid rain

Six‐month‐old Japanese black pine seedlings (Pinus thunbergii) were exposed to simulated acid rain (SAR) at pH 3 and 2 three times a week. After treatment for 2 months, the seedlings were inoculated with a virulent isolate (S10) of the pinewood nematode (Bursaphelenchusxylophilus), at three inoculum levels (P_i = 50, 160 or 500 nematodes per seedling). In seedlings inoculated with 500 nematodes, both population growth of nematodes and disease development were accelerated by pretreatment with SAR at pH 3 or 2. In seedlings inoculated with 50 nematodes, population growth of the nematodes was suppressed and more time was needed for seedlings to die when pretreated with pH 3 SAR. This suggests that exposure to pH 3 SAR increased not only the progress of mortality, but also simultaneously enhanced the tolerance limit of the seedlings to the pinewood nematode – the critical value of physiological burden (represented as a product of time and initial nematode population) necessary to kill a seedling. Exposure to pH 2 SAR accelerated nematode reproduction in seedlings and increased seedling mortality irrespective of the number of nematodes inoculated.     

Role of asymptomatic carrier trees in epidemic spread of pine wilt disease

To determine why pine wilt disease caused by the pinewood nematode (Bursaphelenchus xylophilus) recurs in the same pine stand even after thorough eradication of dead pine trees, the amount of oleoresin exudation from artificial wounds was measured from 72 Pinus koraiensis trees, highly susceptible to this disease, for 4 years. The amount of exuded oleoresin was rated from 0 to 4. All values obtained for each tree were summed at each measurement; thereby a cumulative curve was drawn to monitor the physiological condition of each test tree. Cumulative curves suggest that some pine trees that died had already been infected in the previous year or earlier and then had survived without any visible symptoms. If cessation of oleoresin exudation delays, and overlaps with activity of Monochamus alternatus, the vector beetle of pinewood nematodes, in the following season, such trees can be referred to as latent carriers or asymptomatic carriers. They could play a significant role as attractants for M. alternatus that could then transmit B. xylophilus to neighboring trees. Behavior of M. alternatus caged with several pine seedlings, only one of which acted as an asymptomatic carrier of B. xylophilus, confirmed this idea. The presence of asymptomatic pine trees, which harbored B. xylophilus nematodes or had reduced annual elongation, near stumps of newly dead trees in the study stand also substantiated this hypothesis.     

Discrimination among host tree species by the ambrosia beetle Platypus quercivorus

The ambrosia beetle Platypus quercivorus is a vector for the fungus that causes Japanese oak wilt, and susceptibility to infestation by P. quercivorus varies by tree species. We postulated that P. quercivorus discriminates among host tree species differing in susceptibility to attack. To test this postulate, we counted the number of flying male beetles (NFM), the number of holes bored by male beetles (NH), and the number of flying female beetles (NFF) per unit area of bark surface in three fagaceous tree species: Quercus crispula (with high susceptibility to infestation) and Q. serrata and Castanea crenata (both with low susceptibility). NFM and NH were used to calculate the proportion of male beetles that bored holes out of those that flew to the tree (PBM). We used generalized additive models to predict NFM, NFF, and PBM. The locations of trees, expressed as x and y coordinates, numbers of weeks after the first male beetle’s flying (WEEK), diameters of trees 130 cm above ground (DBH), and tree species (SP) were incorporated into the models as candidate explanatory variables. The best-fit models for NFM and NFF included WEEK and DBH and the effect of location; SP was not included in the models. For PBM, the best-fit model included WEEK, DBH, and SP. The results indicate that male P. quercivorus prefer Q. crispula to Q. serrata and C. crenata and that selection is made before boring holes on trees, but that P. quercivorus do not discriminate among host species when they fly to trees.     

野鸢尾种子至种苗的发育形态学

追踪观察了野鸢尾从种子到种苗的发育过程。结果发现,种子胚苗端发育先于根端;萌发时首先出现下胚轴,继而根环处膨大产生下胚轴毛。种子胚时期,胚芽呈三角形,萌动一段时间后变为舌形,继而胚芽位置开始膨大,向外一侧的子叶鞘逐渐变薄,胚芽突破子叶鞘进入空气之中。野鸢尾可能是由原始水生植物进化来的,因为其种苗发育具有原始水生大型植物的特点。     

A new mycorrhizal helper bacterium, <Emphasis Type="Italic">Ralstonia</Emphasis> species,in the ectomycorrhizal symbiosis between <Emphasis Type="Italic">Pinus thunbergii</Emphasis> and <Emphasis Type="Italic">Suillus granulatus</Emphasis>

In a Robinia-pseudoacacia-dominated coastal forest in Tottori prefecture Japan, the growth and survival of Pinus thunbergii seedlings and the natural regeneration of P. thunbergii was disturbed by R. pseudoacacia. In order to improve the growth of P. thunbergii seedling in the Tottori sand dune, we tried to find a mycorrhiza helper bacteria (MHB) from P. thunbergii mycorrhizosphere in a Tottori sand dune. Two MHB, Ralstonia sp. and Bacillus subtilis, were selected from the nine bacterial species isolated from the mycorrhizosphere of P. thunbergii. The bacterial effect on the ectomycorrhizal fungus Suillus granulatus was investigated by confrontation assay and a microcosm experiment. The confrontation assay showed that Ralstonia sp. promoted the hyphal growth of S. granulatus. Moreover, the S. granulatus–P. thunbergii symbiosis was significantly stimulated by Ralstonia sp. and B. subtilis. Ralstonia sp. and B. subtilis were regarded as MHB associated with P. thunbergii. This is the first report of Ralstonia sp. as an MHB.